KR100530318B1 - Composition comprising papyriflavonol A for control of microorganism - Google Patents

Abstract

The present invention relates to an antimicrobial composition having papyriflavonol A as an active ingredient.

Papyriflavonol A of the present invention exhibits strong antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria, anaerobic acne bacteria and cutaneous candidiasis, and therefore, bacterial skin infections such as acne and atopic dermatitis, and athlete's foot, ringworm, completeness, systemic It can be usefully used for the prevention and treatment of fungal skin infections such as infections and candidiasis.

Description

Composition comprising papyriflavonol A for control of microorganism

The present invention relates to an antimicrobial composition having papyriflavonol A as an active ingredient.

Various skin diseases occurring in the human body are mostly caused by certain pathogenic microorganisms, and are classified into bacterial infections such as acne and atopic disease and fungal infections such as athlete's foot, ringworm, scab, and systemic infection according to the type of infecting microorganism. .

These skin diseases vary greatly in the area and extent of their development, especially opportunistic and systemic infections that can be life-threatening, including long-term use of anticancer agents, the use of immunosuppressants following organ and bone marrow transplantation, and immunity from HIV infection. Due to the deterioration of the body's immune system due to the increase of deficiency patients and the aging of the population, the situation is gradually increasing.

The most common fungal infection is Candida albicans Candidiasis caused by infection is very frequently found in various areas of skin, mouth, pharynx, etc. in patients receiving radiation therapy, pregnant diabetics, burn patients, and children with relatively weak immune function. More than 50% of skin fungal infections are known as candidiasis.

Antimicrobial agents such as erythromycin and chemical fungicides such as azelaic acid are mainly used for the treatment of skin infections that have been developed and used to date. In particular, polyene antibiotics (amphotericin B, nystatin, natamycin), azole antibiotics (fluconazole, ketoconazole, itraconazole), lipopeptide compounds (cilofungin), chitin synthesis inhibitors (polyoxine, nikomycin), nucleic acid analogues (5-fluorocytosine) and the like have been reported. However, due to the widespread use of antibiotics, frequent emergence of resistant strains, irregular absorption, pharmacokinetic vulnerabilities, and low specificity of chemical fungicides, the development of new skin infection treatments that can be used for various pathogenic microorganisms with minimal side effects is urgently needed. . Furthermore, since antifungal agents target eukaryotic cells that have very similar cell physiological characteristics to human cells, they are difficult to show high selective toxicity, resulting in severe cytotoxicity. There are many known side effects, so it is urgent to develop new antifungal treatments to compensate for this.

Therefore, the new antimicrobial therapeutic agent should be 1) safe as an active substance of edible or medicinal plant, and have no side effects due to no side effects, 2) similar to existing antimicrobial agents, or have excellent antimicrobial activity, 3) Consideration should be given to the wide range of activity available for a wide range of patients, 4) to have structural properties that can be easily penetrated into body fluids and tissues, and 5) to facilitate the establishment of rapid mass production systems.

On the other hand, the locust tree (Broussonetia papyrifera Ventenat) is a deciduous broad-leaved small arborescent of the dicotyledonous nettle mulberry family, mainly distributed in Asian tropics and the tropics. The bark is made of various kinds of Korean paper as a raw material of paper, and the fruit is used as a medicine or tastes good, and the young leaves are eaten. It is reported that bast fiber of Coconut tree is used as a raw material of paper, and that various components extracted from Coriander tree have various effects. Specifically, Coconut tree contains tonic, Myeongbok (明目) effect, tone, species, ripening, diuretic, etc., the component that can remove the wind and the component that clears the blood and shows the effect on vision loss, etc. It is reported that it is used as a medicine for physical weakness, decreased vision, and water species in oriental medicine, and it is used as a traditional medicine for species and ascites in China and Indochina.

Accordingly, the present inventors have attempted to search, isolate and purify antimicrobial components of domestic and foreign medicinal and edible plants to develop new skin disease therapeutic agents that can replace or supplement existing antibacterial and antifungal therapeutic agents from natural products that have already been stabilized. In the papyriflavonol A extracted, separated and purified from Root bark of the tree, it was confirmed that there is excellent antibacterial activity against various skin infection bacteria, acne and fungi.

The present invention is to provide an antimicrobial composition comprising papyriflavonol A as an active ingredient.

The present invention provides an antimicrobial composition comprising papyriflavonol A (5,7,3 ', 4'-tetrahydroxy-6,5'-diprenylflavonol) represented by Formula 1 as an active ingredient.

Hereinafter, the present invention will be described in detail.

Papyriflavonol A used in the present invention was used by extracting, separating and purifying according to the preparation method of the Koji tree extract described in Republic of Korea Patent Publication No. 10-361090, the preferred method is as follows.

After cutting 1 kg of dry coriander root bark, the mixture was extracted three times for 5 hours with 1,500 ml of methanol at 65 ° C. using a reflux condenser. The obtained methanol extract was filtered with a funnel using filter paper, reduced pressure and concentrated to obtain 140 g of methanol extract.

140 g of methanol extract was suspended in 1,000 ml of distilled water, and then fractionated three times in a separatory funnel with 1,000 ml of ethyl acetate. The ethyl acetate layer was concentrated to obtain 30 g of ethyl acetate fraction.

30 g of ethyl acetate fraction was subjected to silica gel column chromatography (90 × 6 cm, Merck 7734 600 g) to elute with gradient in the order of chloroform: methanol = 99: 1 to 98: 2 to obtain 7 fractions. The 2nd fraction is separated into two fractions with a solvent of hexane: ethyl acetate = 8: 5 (2-1, 2-2).

Fraction 2-2 was subjected to silica gel column chromatography to elute with gradient in the order of chloroform: methanol = 99: 1 to 98.5: 1.5, and separated into seven small fractions. The small fractions of No. 2 (2-2-2) and No. 3 (2-2-3) were combined, eluted with a solvent of methanol in a Sephadex LH-20 column, and the obtained prillated flavonoid fraction was recrystallized with methanol to give yellow color. Obtain 1 g of papyflavonol A, a pure crystal of needles.

Papyriflavonol A used in the present invention can be obtained not only by being separated and purified by the above method, but also by all conventional methods.

Papyflavonol A of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. The inorganic acid and organic acid may be used as the free acid, and hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, etc. may be used as the inorganic acid, and citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, Fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, galluxuronic acid, embonic acid, glutamic acid or aspartic acid Etc. can be used.

In order to investigate the antimicrobial activity of papyriflavonol A against skin-infected fungi, Candida albicans (KCTC 1940) isolated from bronchial fungal infections and alcohol-fermented yeast Saccharomyces servi known as non-pathogenicity Saccharomyces cerevisiae (IFO 0233) is used.

In addition, to examine the antimicrobial activity against common bacterial infectious diseases, Escherichia coli (KCTC 1924), Salmonella typhimurium (KCTC 1926), Staphylococcus epidermis (ATCC 12228), Staphylococcus aureus (KCTC 1621) is used.

In addition, in order to examine the antibacterial activity against acne bacteria which are chronic skin diseases, propionibacterium acnes (ATCC 6919), which is a chronic skin acne bacterium, is used.

The minimum growth inhibitory concentration for fungi of papyriflavonol A of the present invention is 12.5-25 µg / ml, the minimum growth inhibition concentration for bacteria is 10-25 µg / ml and the minimum growth inhibition for anaerobic acne bacteria is 11 mm. This shows excellent antimicrobial activity against all bacteria.

Therefore, the composition of the present invention can be effectively used for the prevention and treatment of bacterial skin infections such as acne, atopic disease, and fungal skin infections such as athlete's foot, ringworm, scab, systemic infection, candidiasis.

The minimum growth inhibitory concentration of papyflavonol A of the present invention for Candida skin infection fungi is 25 µg / ml, and the minimum growth inhibition concentration for Saccharomyces fermented yeast is 12.5 µg / ml, which completely inhibits growth for 40 hours. do.

In addition, when the concentration of papyriflavonol A of the present invention is increased to 50 µg / ml, which is at least the growth inhibition concentration, it exhibits a marked decrease in absorbance in Candida skin infection fungi and Saccharomyces fermented yeast. In addition, as the treatment concentration of papyriflavonol A of the present invention increases, the cell surface structure is severely destroyed. This is thought to be a decrease in absorbance due to strain dissolution, because papyriflavonol A does not absorb at 600 nm to measure strain growth.

In addition, when the papyriflavonol A of the present invention is treated at various concentrations in Candida skin infection fungi and Saccharomyces fermented yeast, the fluidity of cell membranes in Candida skin infections and Saccharomyces fermented yeast as the concentration increases DPH, which is a fluorescent substance confirming the degree of over-dissolution, releases and releases DPH from the cell membrane to which DPH is bound, thereby decreasing the fluorescent activity of DPH and seriously damaging the cell membrane. In particular, Saccharomyces fermented yeast shows more serious damage below the minimum growth inhibition concentration (12.5 μg / ml).

Therefore, as the concentration of papyriflavonol A of the present invention increases, the absorbance decreases, and the cell surface structure of cell walls, cell membranes, etc. of the fungus of interest is destroyed.

The composition of the present invention may contain one or more active ingredients exhibiting the same or similar functions in addition to the papyriflavonol A.

The composition of the present invention may be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, if necessary, as antioxidants, buffers And other conventional additives such as bacteriostatic agents can be added. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA.

The compositions of the present invention can be administered parenterally (eg, intravenously, subcutaneously, intraperitoneally or topically) or orally, according to the desired method, and topical administration is preferred in the present invention, ointments, creams, emulsions , Plasters, powders, impregnation pads, solutions, gels, sprays, lotions or suspensions.

Dosage varies depending on the weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion and severity of the patient. The daily dose is about 2-6 mg / kg papyflavonol A, preferably 3-4 mg / kg, and more preferably administered once or several times a day.

The composition of the present invention can be used alone or in combination with methods using surgery, hormonal therapy, drug therapy and biological response modifiers for the prevention and treatment of various skin infections.

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

Example 1  : Comparison of antifungal, antibacterial and anti acne activity of papyriflavonol A according to the present invention

In order to investigate the antimicrobial activity of various fungal infections, bacterial infections, and anaerobic bacterial infections of papyriflavonol A according to the present invention, the following experiments were performed using microdilution.

1) antifungal activity

To investigate the antimicrobial activity against skin-infected fungi, Candida albicans (KCTC 1940) isolated from bronchial fungal infections and Saccharomyces cerevisiae (IFO 0233) known as non-pathogenic pathogens were used. Used.

These strains were subcultured every four weeks using Potato Dextrose Agar (Difco. Co), and the culture of seed strains was based on Sobraud-glucose medium (Sabouraud dextrose broth; bactopeptone 1%, dextrose 1%). Used.

As a control for evaluating the antifungal activity of papyriflavonol A sample, amphotericin B, miconazole and 5-fluorocytosine, which are used as antifungal agents, were used as sigma. Co. St. Louis, MO) was used, and was dissolved in dimethylsulfoxide (dimethylsulfoxide) in the same manner as papyriflavonol A and diluted to an appropriate concentration.

Candida albicans and Saccharomyces cervicia were pre-incubated in 5 ml of Sobrawood-glucose medium at 30 ° C. for 8 hours, and the control group (amphotericin vi, myconazole, 5-fluorocytosine) at a constant concentration. And 0.3 ml of a dilution of Papyflavonol A sample was mixed with 2.7 ml of Sobrid-Glucose Medium.

Thereafter, each seed was inoculated to a concentration of 1 × 10 ⁵ CFU / mL, and then incubated at 28 ° C. for 24 hours to determine the minimum concentration of the sample without bacterial growth as the minimum growth inhibition concentration (MIC). It was. All experiments were repeated three or more times.

The results are shown in Table 1.

2) antibacterial activity

To investigate the antimicrobial activity against common bacterial infectious diseases, Escherichia coli (KCTC 1924), Salmonella typhimurium (KCTC 1926), Staphylococcus epidermis (ATCC 12228), Staphylo Staphylococcus aureus (KCTC 1621) was used.

These strains were subcultured every two weeks using nutrient solid medium (Nutrient Agar, Difco. Co), and the culture of the seed strain was used nutrient liquid medium (Nutrient broth, Difco. Co).

As a control for evaluating antibacterial activity of Papyriflavonol A sample, ampicillin and erythromycin were purchased from Sigma Co., Ltd. (Sigma Co. St. Louis, MO) at a concentration suitable for sterile distilled water. It was used by melting.

Pure colonies of Escherichia coli, Salmonella typhimurium, Staphylococcus epidermis and Staphylococcus aureus were preincubated for 8 hours at 37 ° C. in 5 ml of Nutrient Broth, Difico. Co. 0.3 ml of the dilutions of the control group (Epicillin, erythromycin) and Papyflavonol A samples of concentration were mixed with 2.7 ml of nutrient solution medium.

Thereafter, each seed was preincubated at a concentration of 1 × 10 ⁵ CFU / mL, and then incubated at 37 ° C. for 24 hours to determine the minimum concentration of the sample without bacterial growth on the naked eye. All experiments were repeated three or more times.

The results are shown in Table 1.

3) Anti acne activity for antibacterial effect of anaerobic bacteria

To investigate the antimicrobial activity of acne bacteria, a chronic skin disease, the prophynibacterium acnes (ATCC 6919), an anaerobic incubator and anaerobic solid medium (anaerobic agar, Difico. Co), was used. 24 hours preculture at 37 ℃, subcultured every two weeks was used.

10 ⁸ bacteria were smeared in an anaerobic solid medium under the same conditions as the passage condition, and 5 mm diameter filter paper (Whatmann Co.) was aseptically placed on the solid medium.

Subsequently, 10 µg of each of the controls (Empicillin, Erythromycin, Ampoterisin B, Myconazole, 5-fluorocytosine) dissolved in dimethyl sulfoxide and 100 µg of Papyriflavonol A sample were added dropwise onto a sterile filter paper. After incubation in an anaerobic incubator for 40 hours, the anti-acne bacterium activity was evaluated by measuring the size of the growth inhibitory rings formed around the sterile filter paper. All of the above operations were performed in an anaerobic culture zone filled with nitrogen only.

The results are shown in Table 1.

Antimicrobial Activity of Papyriflavonol A against Antifungal, Antibacterial and Antiacne Antimicrobial activity Minimum growth inhibitory concentration (MIC; μg / ml) Growth support ring (mm) Antifungal Antibacterial Anti acne Candida albicans Saccharomyces Serbia Escherichia coli Salmonella typhimurium Staphylococcus Epidermis Staphylococcus aureus Propionibacterium Acnes Papyriflavonol A 25 12.5 20 25 10 15 11 Empicillin - - 1.25 1.25 20 1.25 70 Erythromycin - - 1.25 5 1.25 1.25 70 Amphotericin vii 1.25 1.25 - - - - - Myconazole 1.25 1.25 - - - - 32 5-fluorocytosine > 50 6.0

As shown in Table 1, the minimum growth inhibitory concentration of papyflavonol A of the present invention is 12.5-25 μg / ml for fungi, 10-25 μg / ml for bacteria, and growth inhibition rate for anaerobic acne bacteria. 11 mm.

In addition, the control groups of empicillin and erythromycin showed no antimicrobial activity against fungi such as Candida and Saccharomyces, and the minimum growth inhibitory concentration of empicillin against various pathogenic bacteria was 1.25-20 ㎍ / ml, erythromycin. The minimum growth inhibition of was 1.25 ~ 5 ㎍ / ㎖, and the growth inhibition rate for anaerobic acne was 70 mm.

In addition, amphotericin B, myconazole and 5-fluorocytosine also showed excellent antimicrobial activity against fungi, but showed no antimicrobial activity to bacteria.

As described above, papyriflavonol A exhibits excellent antimicrobial activity against all bacteria including fungi, bacteria and anaerobic acne bacteria, so that the composition of the present invention can be effectively used for the prevention and treatment of various skin infection microorganisms.

Example 2  : Minimal growth inhibitory concentration of papyriflavonol A according to the present invention against fungi (candida skin infection fungi and Saccharomyces fermented yeast)

In order to determine the minimum growth inhibitory concentration of the fungi of papyriflavonol A according to the present invention, the change of the growth of the fungus was confirmed by spectroscopic method while treating the fungus with papyriflavonol A for 40 hours.

Candida and Saccharomyces strains were pre-incubated in 5 ml of nutrient liquid medium (Nutrient Broth, Difco, Co) at 30 ° C. for 8 hours, and then 0.1 ml of the pre-culture solution was dissolved in dimethyl sulfoxide in papyriflavone A. After addition to 2.9 ml of a new nutrient liquid medium containing 25 µg / ml or 12.5 µg / ml, respectively, the growth of Candida and Saccharomyces strains was measured by spectroscopic method at 600 nm while incubating at 30 ° C. Asys-HITECH, Expert 96, Asys Co. Austria).

As a control, only 2.9 ml of dimethyl sulfoxide was added to 2.9 ml of new nutrient liquid medium, and the measurement was performed in the same manner as described above.

The results are shown in FIG.

As shown in Fig. 1, the concentration of papyriflavonol A was 25 µg / ml in Candida fungi and 12.5 µg / ml in Saccharomyces fermented yeast, which completely inhibited growth for 40 hours. A decrease in absorbance was observed.

Example 3  : Minimum growth inhibitory concentration for fungi (Candida skin infection fungi and Saccharomyces fermented yeast) with increasing concentration of papyriflavonol A according to the present invention

In order to observe the growth rate that occurs when the concentration of papyriflavonol A of the present invention is treated above the minimum growth inhibition concentration, it was measured by the following spectroscopic method.

Candida fungi and Saccharomyces fermented yeast were treated with the concentrations of 25 μg / ml and 50 μg / ml of papyflavonol A and measured by the same spectroscopic method as in Example 2.

As a control, 0.1 ml of dimethyl sulfoxide was used to test the effect of dimethyl sulfoxide.

The results are shown in FIG.

As shown in FIG. 2, when papyriflavonol A was treated with 25 μg / ml and 50 μg / ml, there was a marked decrease in cell absorbance against Candida fungi and Saccharomyces fermented yeast. On the other hand, the cell absorbance increased in all the controls.

Example 4  : Cell lysis electron micrograph of Candida skin infection fungi and Saccharomyces fermented yeast according to various concentrations of papyriflavonol A of the present invention

The change of cell surface structure according to various concentrations of papyriflavonol A of the present invention was observed by electron microscopy.

Candida skin infection fungi and Saccharomyces fermented yeast at the beginning of the logarithmic phase were each suspended in phosphate buffer (pH 7.4) at 10 ⁸ CFU / ml, followed by various concentrations of papyflavonol A (0 µg / ml, 12.5 μg / ml, 25 μg / ml, 50 μg / ml) was added and incubated at 30 ° C. for 2 hours. Thereafter, Na-cacodylate buffer solution containing 5% glutaraldehyde (0.2M, pH 7.4) was added in the same volume to fix the cells at 4 ° C. for 2 hours.

Thereafter, fixed cells were recovered using an isophore filter (Isopore filters, 0.2 μm pore size, Millipore, Bedford, Mass.) And washed with excess Na-cacodylate buffer to remove glutaraldehyde.

Each washed cell was treated with 1% osmium tetroxide, washed with Na-cacodylate buffer containing 5% sugar, and gradually dehydrated using various concentrations of ethanol. Finally, after the critical drying, gold coating was performed, and the cell surface structure was observed by magnification 10,000 times with a scanning electron microscope (Hitachi, S-2500C, Japan).

As a control group, each subject fungus was treated with dimethyl sulfoxide alone, and then fixed, washed, dehydrated, and applied in the same manner, and observed with a scanning electron microscope.

The results are shown in FIG.

As shown in FIG. 3, the cell surface structure of the Candida strain or Saccharomyces strain in the control group treated with dimethyl sulfoxide alone remained stable. On the other hand, when treated with papyriflavonol A, as the treatment concentration increased, the cell surface structure was severely destroyed in a concentration-dependent manner.

Therefore, it can be seen that as the concentration of papyriflavonol A increases, it causes severe damage to the superficial structure of the target cell and inhibits growth.

Example 5  : Measurement of Cell Membrane Destruction of Fungi by Target Treatment of Papyriflavonol A of the Present Invention

In order to determine whether papyriflavonol A of the present invention induces dynamic fluidity and breakage of the cell membrane, the following experiment was performed.

Cells of Candida skin-infected fungi or Saccharomyces fermented yeast were pre-incubated for 8 hours at 30 ° C. in nutrient liquid medium (Nutrient Broth, Difco Co.) to recover the cells of the logarithmic growth phase. The recovered cells were added to a fresh nutrient liquid medium at 10 ⁸ CFU / mL, and various concentrations of papyriflavonol A were added, followed by further incubation at 30 ° C for 3 hours.

Dimethyl sulfoxide was added as a control.

The treated cells were fixed at 30 ° C. for 30 minutes at 0.25% formaldehyde and then collected by centrifugation at 3000 rpm for 5 minutes. Aggregated cells were washed again three times with phosphate buffer (pH 7.4) and recovered, soaked in liquid nitrogen and frozen.

Afterwards, the frozen cells were re-dissolved in phosphate buffer (pH 7.4) to determine the degree of cell membrane damage, and DPH (1,6-diphenyl-1,3,5-hexatriene; Molecular Probes, Inc., Eugene, OR, USA) ) Was added to a final concentration of 10 ⁻⁷ M and reacted at 30 ° C. for 45 minutes. Thereafter, unreacted DPH was removed by washing with excess phosphate buffer (pH 7.4) five times, and then the amount of DPH bound to the cells was measured using a F-4500 fluorescence spectrometer (Fluorescence Spectrophotometer, Hitachi, Tokyo, Japan). . In the measurement, the excitation wavelength was fixed at 350 nm and the emission wavelength at 425 nm.

The results are shown in Table 2.

Cell membrane-bound DPH fluorescence activity by papyriflavonol A treatment (%) Papyriflavonol A treatment concentration (µg / ml) Cell membrane-bound DPH fluorescence activity (%) Candida skin fungus Saccharomyces fermented yeast 0 100.0 100.0 3.125 85.7 59.6 6.25 53.5 34.2 12.5 64.0 21.7 25.0 36.5 20.3

As shown in Table 2, fluorescence activity of DPH was induced by the release of DPH from the DPH-coupled cell membrane in Candida skin-infected fungi and Saccharomyces fermented yeast as the concentration increased during the treatment of papyriflavonol A. This was reduced, and cell membrane damage appeared seriously. Saccharomyces fermented yeast also showed more severe damage below the minimum growth inhibition concentration (12.5 μg / ml).

Experimental Example  : Oral Acute Toxicity Test

In order to confirm whether papyflavonol A of the present invention is toxic in vivo, an acute toxicity test was carried out in the following manner.

Six-week-old SPF rats were divided into five rats per group, and the papyflavonol A of the present invention was suspended in 0.5% methylcellulose solution and administered orally at a dose of 300 mg / kg. After administration, mortality, clinical symptoms, and changes in body weight were observed, and hematological and blood biochemical tests were performed. Necropsy was performed to observe abdominal and thoracic organ abnormalities.

As a result, there were no clinical symptoms or dead animals in the animals administered papyflavonol A of the present invention, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, autopsy findings, and the like. The papyriflavonol A used in the experiment did not show a toxicity change up to 300 mg / kg in rats, and the minimum lethal dose (LD ₅₀ ) upon oral administration was determined to be 300 mg / kg or more.

Examples of pharmaceutical formulations for the compositions of the present invention are illustrated below.

Formulation Example 1  : Manufacturing method of ointment

Papiliflavonol A 5.0 mg

Stearic acid 5.5 mg

Cetyl Alcohol 150 mg

50 mg of liquid paraffin

Polyoxylstearate # 40 40 mg

Ethyl paraben 0.950 mg

Glycerin 5.5 mg

Purified water

The above components were mixed and prepared by stirring and dispersing by a conventional ointment manufacturing method.

Formulation Example 2  : Manufacturing method of spray

Papiliflavonol A 5.0 mg

Phloxamer 188 100.0 mg

Methyl paraoxybenzoate 2.0 mg

Sodium dihydrogen phosphate 3.4 mg

Sodium chloride 8.3 mg

Phosphoric acid

Purified water

The above components were mixed and prepared by stirring and dispersing by the usual spray production method.

Formulation Example 3  : Manufacturing method of injection solution

Papyflavonol A 0.6 g

0.1 g sodium chloride

Distilled water

After mixing the above components, the mixed solution was filled into a 5 ml type I ampoule made of transparent glass, and the glass was dissolved under the upper lattice of air by dissolving the glass. Was prepared.

Formulation Example 4  : Manufacturing method of syrup

Papyflavonol A 2g

Saccharin 0.8 g

25.4 g per

Glycerin 8.0 g

0.04 g of spices

Ethanol 4.0 g

0.4 g of sorbic acid

Distilled water

After mixing the above components, it was prepared according to the conventional method for producing a syrup.

Formulation Example 5  : Manufacturing method of tablet

Papiliflavonol 250 g

Lactose 175.9g

Potato Starch 180g

32g colloidal silicic acid

10% gelatin solution

50 g of talc

5 g magnesium stearate

After mixing the above components, it was prepared by tableting according to the conventional method for producing tablets.

Papyriflavonol A, which is extracted, separated and purified from the Root bark of Coriander according to the present invention, is a variety of Gram-positive bacteria, unlike conventional antibiotics having a limited range of antimicrobial activity that can control only bacteria or fungi. Excellent antimicrobial activity against gram negative bacteria, anaerobic acne bacteria and cutaneous candidiasis fungi.

Therefore, the composition of the present invention can be usefully used for the prevention and treatment of bacterial skin infections such as acne, atopic disease, and fungal skin infections such as athlete's foot, ringworm, striation, systemic infection, candidiasis.

1 is a diagram showing a minimum growth inhibitory effect when the concentration of papyriflavonol A of the present invention was treated with Candida fungi at 25 µg / ml and Saccharomyces fermented yeast at 12.5 µg / ml.

Figure 2 shows the effect of growth inhibition on Candida fungi and Saccharomyces fermented yeast when papyriflavonol A of the present invention was treated at a minimum growth inhibitory concentration (25 µg / ml, and 50 µg / ml). It is also.

3 is an electron microscope observation of changes in the cell surface structure of Candida fungi and Saccharomyces fermentation yeast according to various concentrations of papyriflavonol A of the present invention.

Claims (3)

An antimicrobial composition comprising papyriflavonol A as an active ingredient. The antimicrobial composition according to claim 1, wherein the papyriflavonol A is extracted, separated, and purified from the beech tree. The antimicrobial composition of claim 1, which is useful for the prevention and treatment of bacterial skin infections such as acne, atopic disease, and fungal skin infections such as athlete's foot, ringworm, rash, systemic infection, candidiasis.